• Title/Summary/Keyword: Tet-On

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Antibiotic resistance pattern of avian pathogenic Escherichia coli isolated from chickens (닭에서 분리된 조류 병원성 대장균의 항생제 내성 양상)

  • Kim, Myeong Suk;Kwon, Hyuk Moo;Sung, Haan Woo
    • Korean Journal of Veterinary Research
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    • v.49 no.3
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    • pp.195-200
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    • 2009
  • Avian pathogenic Escherichia coli (APEC) is an important bacterial pathogen of chickens and causes colibacillosis such as airsacculitis, perihepatitis, omphalitis, peritonitis, salpingitis, and pericarditis. As the transfer of antibiotic resistance from animal to humans can be possible, surveillance on antibiotic resistance of APEC is very important. A total 34 APEC isolates from diseased chickens during the period from 2007 to 2009 were obtained. The susceptibility of the isolates to 13 antibiotics was determined by disc diffusion assay. Resistance to erythromycin was found in 97.1% of APEC isolated, followed by resistance to tetracycline (85.3%), doxycycline (82.3%), ampicillin (73.5%), sulfisoxazole (67.6%), enrofloxacin (67.6%), ciprofloxacin (64.7%), norfloxacin (61.7%) trimethoprim/sulfamethoxazole (52.9%), gentamycin (26.5%), amoxicillin (8.8%), colistin (5.9%), and amikacin (2.9%). The blaTEM genes were detected in 25 (100%) of the 25 ampicillin-resistant APEC isolates. Among the 29 tetracycline-resistant APEC isolates, tetA and tetB genes were detected in 18 (62.1%) and 9 (31%) isolates, respectively. Twenty six (76.5%) isolates were multiresistant to at least 6 antibiotics and seven (20.1%) isolates were multiresistant to at least 10 antibiotics. This results indicated that multiple antibiotic-resistant APEC is widespread in chicken flocks in Korea.

Production of hTPO Transgenic Chickens using Tetracycline-Inducible Expression System (Tetracycline-Inducible Expression System을 이용한 Human Thrombopoietin (hTPO) 형질전환 닭의 생산)

  • Kwon, M.S.;Koo, B.C.;Kim, D.H.;Kim, M.J.;Kim, T.
    • Korean Journal of Poultry Science
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    • v.36 no.2
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    • pp.177-186
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    • 2009
  • It is well-known that unregulated over-expression of foreign gene may have unwanted physiological or toxic effects in transgenic animals. To circumvent these problems, we constructed retrovirus vector designed to express the foreign gene under the control of the tetracycline-inducible promoter. However, gene expressions in the tetracycline-inducible expression system (Tet system) are not completely regulated but a little leaky due to the inherent defects in conventional Tet-based systems. A more tightly controllable regulatory system can be achieved when the advanced versions ($rtTA2^SM2$) of rtTA and a minimal promoter in responsive components (pTRE-tight) are used in combination therein. In this study, we tried to produce human thrombopoietin (hTPO) from various target cells and transgenic chickens using the retrovirus vector combined with Tet system. hTPO is the primary regulator of platelet production and has an important role in the survival and expansion of hematopoietic stem cells. In a preliminary experiment in vitro, higher hTPO expression and tighter expression control were observed in chicken embryonic fibroblast (CEF) cells. We also measured the biological activity of the hTPO using Mo7e cells whose proliferation is dependant on hTPO. The biological activity of the recombinant hTPO from CEF was higher than both its commercial counterpart and hTPO from other target cells. The recombinant retrovirus was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 138 injected eggs, 15 chicks hatched after 21 days of incubation. Among them, 8 hatched chicks were hTPO positive. When the Go transgenic chicken was fed doxycycline (0.5 mg per 1 gram of feed), a tetracycline derivative, hTPO concentration of the transgenic chicken blood was 200 ng/mL. Germline transmission of the transgene was confirmed in sperm of the Go transgenic roosters. These results are informative to establish transgenic chickens as bioreactors for the mass production of commercially valuable and biological active human cytokine proteins.

Retrovirus Vector-Mediated Construction of Porcine Cell Lines Expressing Human PTH (Retrovirus Vector System을 이용한 hPTH가 발현되는 돼지 세포의 구축)

  • Jeong J. Y.;Koo B. C.;Kim N. H.;Kwon M. S.;Park S. H.;Kim T. O.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.197-202
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    • 2004
  • Human PTH (parathyroid hormone) is known to be efficacious for curing osteoporesis. In this study, we attempted to construct genetically modified porcine cell lines that can ultimately be used for donor cells in nuclear transfer-mediated transgenesis. By using retrovirus vectors carrying tetracycline-regulatory expression system and WPRE (woodchuck hepatitis virus posttranscriptional regulatory element) sequence, we could control PTH expression with tetracycline and boost the promoter activity. Considering that low or constitutive expression of the transgene has been one of major problems that needs to be solved in transgenic animal studies, our results could be helpful in successful production of transgenic pigs as bioreactors.

Expression of the Recombinant Porcine GH Gene In Vitro Using Tetracycline Inducible Expression System (In Vitro에서의 Tetracycline Inducible Expression System에 의한 재조합 돼지 성장호르몬 유전자의 발현)

  • Kwon Mo Sun;Koo Bon Chul;Kim Teoan
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.49-55
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    • 2005
  • We cloned cDNA of the PGH(porcine growth hormone) gene and constructed retrovirus vector designed to express PGH gene under the regulation of CMV (cytomegalovirus) promoter. To maximize the expression, WPRE(woodchuck hepatitis virus posttranscriptional regulatory element) sequence was placed at the downstream of the PGH gene. After infection with recombinant viruses, approximately 1×10/sup 6/ PFF(porcine fetal fibroblast) cells released PGH protein into the media as much as 1,400 ng. In a subsequent experiment, a modifications of the retrovirus vector was made to express the PGH gene in a teracycline-inducible manner. In PFF cells carrying these viral vector sequences, addition of doxycycline to the media resulted in 2∼6 fold increase in PGH synthesis. In the modified retrovirus vectors, the WPRE sequence also played a role in boosting the effect of the tetracycline induction. This result indicates that our tetracycline-inducible expression system might be a promising candidate in alleviating the complicate physiological problems caused by constitutive expression of the exogenous genes in the transgenic animals.

Antimicrobial Resistance and Implicated Genes of E. coli Isolated from Commercial and Cooked Foods in Seoul (서울시내 유통식품에서 분리한 대장균의 항생제 내성 및 내성유전자)

  • Yoo, Young-Ah;Kim, Moo-Sang;Kim, Kyong-Sik;Park, Sun-Hee;Jung, Sung-Kuk
    • Journal of Food Hygiene and Safety
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    • v.25 no.3
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    • pp.220-225
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    • 2010
  • Distribution of foodborne E. coli strains, antimicrobial resistant genes and antimicrobial susceptibility have been carried out on E. coli isolated from commercial and cooked foods distributed food in Seoul. Of total 1,313 samples, fifty samples(3.8%) were found E. coli that included one of the ETEC and EPEC, respectively. The serotype of ETEC in seasoning raw meat was E. coli O26 and produced Verotoxin 2. Fifty percentage of total isolates were susceptible to all antimicrobial agents. Specially, there were ampicillin(36%), amoxicillin/clavulanic acid(32%) and tetracycline(22%) etc. Resistant gene (tetB) were found in four tetracycline resistant E. coli strains, and TEM gene was found in one ampicillin resistant E. coli isolate.

Construction of Retrovirus Vector System for the Regulation of Recombinant hTPO Gene Expression (재조합 hTPO 유전자의 발현 조절을 위한 Retrovirus Vector System의 구축)

  • Kwon, Mo-Sun;Koo, Bon-Chul;Kim, Do-Hyang;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.161-167
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    • 2007
  • In this study, we constructed and tested retrovirus vectors designed to express the human thrombopoietin gene under the control of the tetracycline-inducible promoters. To increase the hTPO gene expression at him-on state, WPRE sequence was also introduced into retrovirus vector at downstream region of either the hTPO gene or the sequence encoding reverse tetracycline-controlled transactivator (rtTA). Primary culture cells (PFF, porcine fetal fibroblast; CEF, chicken embryonic fibroblast) infected with the recombinant retrovirus were cultured in the medium supplemented with or without doxycycline for 48hr, and induction efficiency was measured by comparing the hTPO gene expression level using RT-PCR, western blot and ELISA. Higher hPTO expression and tighter expression control were observed from the vector in which the WPRE sequence was placed at downstream of the hTPO (in CEF) or rtTA(in PFF) gene. This resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.

Education and Professional Development of Mathematics Teachers in Korea

  • Park, Han-Shick;Shin, Hyunyong
    • Research in Mathematical Education
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    • v.8 no.2
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    • pp.69-80
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    • 2004
  • It is undeniable that teachers play the principal roles in education. This is why education and professional development of teachers are so important. Some of recent works have made this fact clearer. In America, in particular, many reports and research papers have recently been published on these problems. In this paper, we first introduce briefly the current system of education, employment, and professional development of mathematics teachers in Korea. And then we mention a research project on education of mathematics teachers. The final report of the project contains some suggestions for curriculums of the department for education of mathematics teachers. We describe one example of extended syllabus which implements those suggestions. The example is on Modern Algebra I.

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Ensiled Green Tea Waste as Partial Replacement for Soybean Meal and Alfalfa Hay in Lactating Cows

  • Kondo, Makoto;Nakano, Masashi;Kaneko, Akemi;Agata, Hirobumi;Kita, Kazumi;Yokota, Hiroomi
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.7
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    • pp.960-966
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    • 2004
  • The purpose of this study was to evaluate the effects of protein supplementation of green tea waste (GTW) on the performance of lactating cows. Another aim was to increase resource utilization and to eliminate any environmental negative impact from the tea waste. GTW from a beverage company was ensiled at a low pH (<4.0) and high acetic acid and lactic acid concentration, and it contained high crude protein (CP, 34.8%), total extractable tannins (TET, 9.2%) and condensed tannin (CT, 1.7%). Two experiments were conducted to investigate the palatability and performance in lactating cows fed GTW. In the palatability trial, three lactating cows were allocated to three dietary treatments in a 3$\times$3 Latin square design. The animals were offered a total mixed ration (TMR) including GTW at rates of 0, 2.5 and 5.0% on a dry matter (DM) basis. Total DM intake was not different among the treatments. In the performance trial, four lactating cows were used in a 2$\times$2 Latin square design with a 3 week sampling period. GTW was incorporated into TMR at a rate of 5.0% on a DM and 10.0% on a CP basis. Thus GTW replaced alfalfa hay and soybean meal at a level of 25.0% on a DM. DM and CP intake were not affected by the inclusion of GTW, whereas TET and CT intake were significantly increased (p<0.001). Milk production, milk composition and the efficiency of milk production were not altered by the GTW inclusion. Although ruminal pH and VFA, and blood urea nitrogen were not changed, ruminal $NH_{3}-N$ and plasma total cholesterol were relatively low in the GTW group, but not significantly different. The excretion of urinary purine derivatives and estimated MN supply were also not significantly affected by GTW treatment. It is therefore concluded that GTW can be used as a protein source without any detrimental effects on the performance of lactating cows.

Studies on the improvement of the productivity of purse seine fishery-III - The characteristics on the motion with the flow velocity of model purse seine of the subjective power block and triplex during pursing - (선망어업의 생산성 향상에 관한 연구-III - 죔줄 체결시 파워불록과 트리플랙스용 선망 모형의 유속에 따른 운동특성 -)

  • Kim, Suk-Jong
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.43 no.1
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    • pp.12-27
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    • 2007
  • This fundamental studies on for the productivity improvement and laborsaving of purse seine fishery. Given the difficulty posed from the distortion of net shape caused by the external forces, such as tide, at the time of shooting and pursing, we set the 4 steps of 0, 2, 4 and 6cm/sec in flow velocity in the flume tank for the experiment in order to examine those characteristics. We used two model seines designed on the scale of 1 to 180 based on the power block seine, which is the mackerel purse seine generally used in the near sea of Jeju Island and triplex seine, which is the mackerel purse seine of one boat system fishing expected in the future, for the experiment, and interpreted the characteristics of several motion in water, such as the shape of seine, the change in tension and area during pursing and its the analysis results are as follows. Though the experiment could be conducted up to 6cm/sec of flow velocity that was defined, the experiment could not go on because of the severe distortion in the seine at the flow velocity in excess of 6cm/sec. As for the depth of leadline and reduction rate of side area of seine when the pursing is connected, P seine turned out to be slightly higher than T seine, and the hauling speed and reduction rate of upper area of seine were found similar to each other. The correlation between the hauling time (Ht) and depth of lead line (Dhp, Dht) of P seine and T seine can be expressed by the equation, that is, Dhp=(0.99Pt-7.63)Pt+69.01, Dht=(1.03Pt-7.73)Pt+66.74. The correlation between the hauling time and hauling velocity (Hpp, Hpt) can be expressed by the equation, that is, $Hpp=-0.06Ht^2+0.88Ht+0.78,\;Hpt=-0.05Ht^2+0.81Ht+0.98$ here, Pt is pursing time. And the correlation between the pursing time and the reduction rate of side area (sArp, sArt) can be expressed by the equation, that is, $sArp=-0.48Pt^2+14.79Pt-16.74,\;sArt=-0.45Pt^2+14.56Pt-16.48$. The reduction rate of upper area of seine (tArp, tArt) can be expressed by the equation, that is, $tArp=0.34Pt^2-0.66Pt-0.74,\;tArt=0.34Pt^2-0.27Pt-1.80$. In addition, the correlation between the pursing time and tension of purse line (Tep, Tet) can be expressed by the equation, that is, $Tep=2.79Pt^2+2.26Pt-0.60,\;Tet=2.14Pt^2+8.08Pt-27.50$.

Detecting DNA hydroxymethylation: exploring its role in genome regulation

  • Sun-Min Lee
    • BMB Reports
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    • v.57 no.3
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    • pp.135-142
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    • 2024
  • DNA methylation is one of the most extensively studied epigenetic regulatory mechanisms, known to play crucial roles in various organisms. It has been implicated in the regulation of gene expression and chromatin changes, ranging from global alterations during cell state transitions to locus-specific modifications. 5-hydroxymethylcytosine (5hmC) is produced by a major oxidation, from 5-methylcytosine (5mC), catalyzed by the ten-eleven translocation (TET) enzymes, and is gradually being recognized for its significant role in genome regulation. With the development of state-of-the-art experimental techniques, it has become possible to detect and distinguish 5mC and 5hmC at base resolution. Various techniques have evolved, encompassing chemical and enzymatic approaches, as well as third-generation sequencing techniques. These advancements have paved the way for a thorough exploration of the role of 5hmC across a diverse array of cell types, from embryonic stem cells (ESCs) to various differentiated cells. This review aims to comprehensively report on recent techniques and discuss the emerging roles of 5hmC.